http://dx.doi.org/10.21577/0103-5053.20200017

J. Braz. Chem. Soc., Vol. 31, No. 6, 1317-1326, 2020 Article Printed in Brazil - ©2020 Sociedade Brasileira de Química

Occurrence of Pesticides Associated to Atmospheric Aerosols: Hazard and Cancer Risk Assessments

Aleinnys M. B. Yera, *,a Madson M. Nascimento,b,c,d Gisele O. da Rocha,b,c,d Jaílson B. de Andradeb,c,e and Pérola C. Vasconcellos *,a,b

aInstituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes, 748, 05508-000 São Paulo-SP, Brazil bInstituto Nacional de Ciências e Tecnologias de Energia e Ambiente (INCT), Universidade Federal da Bahia, Campus Ondina, 40110-903 Salvador-BA, Brazil cCentro Interdisciplinar de Energia e Ambiente (CIEnAm), Universidade Federal da Bahia, Campus Ondina, 40110-903 Salvador-BA, Brazil dInstituto de Química, Universidade Federal da Bahia, Campus Ondina, 40110-903 Salvador-BA, Brazil eSENAI-CIMATEC Centro Universitário, 41650-110 Salvador-BA, Brazil

Since the 1950s, pesticides have been used in agriculture. The increase in their consumption has been observed in recent years. In this work, it was determined the concentration of pesticides in the atmosphere of São Paulo and Piracicaba cities (sugarcane plantation site) and the hazard quotients and cancer risk caused by inhalation of these compounds were calculated. Twenty‑three samples were analyzed and 34 pesticides associated to atmospheric particulate

matter (with diameter equal to or less than 2.5 (PM2.5) and 10 µm (PM10)) were investigated by gas chromatograpy-mass spectrometry (GC-MS). Fourteen compounds including organochlorines, organophosphorus, and pyrethroids pesticides, were determined in these samples. The concentrations in the particulate matter ranged from 17 pg m−3 (tebuconazole) in Piracicaba, to 166 pg m−3 (endrin aldehyde) in São Paulo. The highest values of daily inhalation exposure for heptachlor were 9.0 × 10−5 mg kg−1 day−1 at Piracicaba, and 6.5 × 10−5 mg kg−1 day−1 at São Paulo, in both cases registered in infants. The values of the hazard quotients are lower than 1.0, indicating that there is no danger to the exposed population, it is health protective. On the other hand, the cancer risk calculations for heptachlor resulted in values above those recommended by US Environmental Protection Agecy (EPA).

Keywords: air pollution, particulate matter, pesticides concentration, risk assessment

Introduction spray drift process. Post-application emission is another way of pesticide input in the atmosphere. The properties of The intensive use of pesticides has been leading to the molecules that most influence on gas-particle distribution contamination of several environmental compartments, are: vapor pressure, Henry’s constant and persistence in including the atmosphere. the air. However, the vapor pressure is considered the key The pesticides distribution in the atmosphere depends factor that governs this partitioning.1 on the pesticides physicochemical properties as well as In the last few years, different types of pesticides have the meteorological conditions of a given study area. In been detected in the atmosphere in rural and urban areas general, the pesticide application can be done by aircraft around the world. Villiot et al.2 evaluated the concentrations or land spraying. During the application, from 30 to 50% of current used pesticides in the center of Reims (France). of the applied amount can be lost in the atmosphere due to A total of 197 samples of particles with diameter equal

to or less than ten micrometers (PM10) were collected for three years (from 2012 to 2015), and the results showed *e-mail: [email protected]; [email protected] 28 pesticides detected.2 1318 Occurrence of Pesticides Associated to Atmospheric Aerosols J. Braz. Chem. Soc.

Another important information is the risk assessment; October 2008, about 3 m above the ground level. The result it presents the exposure risk of human to different types obtained in these samples will allow us to perform a brief of pesticides. It can occur via dermal, oral, and inhalation initial characterization of the pesticides behavior in the exposures, the latter reaching two or three orders of atmosphere of the rural site. magnitude smaller than oral exposure.1 Even so, the The other city studied was São Paulo (SPA). The inhalation exposure is important. Viel et al.3 associated metropolis is the capital of São Paulo State and most the presence of organochlorine pesticides in the air with populous city in Brazil, with over 12 million inhabitants the risk of developing non-Hodgkin lymphoma (NHL) (Figure S2, SI section). SPA have a strong industrial in a population exposed to pesticides.3 Due to massive character. Agriculture is not an important sector of its and indiscriminate use of these compounds, the human economy, only 7% (11 thousand ha) of its total area is population, especially infants and children, have presented dedicated to this activity, being beans and cassava its chronic effects such as the loss of coordination and memory principal crops.9 Pesticides as azoxystrobin, bifenthrin, related to Parkinson’s and Alzheimer’s diseases.4 In this carbofuran, λ-cyhalothrin, and permethrin used in beans way, evaluations of daily inhalation exposure and hazard and cassava crops, gardens, and the domestic pest control quotients in all populations are indispensable to assess the are the most consumed.10 risk of the pesticides. Samples of particles with diameter equal to or less than

In 2017 the consumption of active products in Brazil 2.5 micrometers (PM2.5) were collected about 20 m above reached 540 thousand tons, locating the country as one the ground level, in the rooftop of a building (Institute of the largest consumers of these worldwide. Currently, of Astronomy, Geophysics and Atmospheric Sciences, Brazilian laws5 permit the use of 526 different active University of São Paulo). The location is inside a green products, out of which 30% of them are banned in the area and approximately 2 km away from an important

European Union and in the United States of America, highway. PM2.5 samples (n = 15) were collected in August including: atrazine and acephate. In 2016, the consumption 2017, with the aim of studying pesticides behavior in the of atrazine reached 29 thousand tons.6 The state of São atmosphere of an urban site. Paulo is the second major consumer of pesticides in Brazil, The samples in both sites were collected using a high- over 110 thousand tons used in 2016.7 volume sampler at flow rate 1.13 3m min−1, with 2.5 and The principal objective of this work was to study the 10 µm size selective inlets (Thermo Andersen, USA). The pesticides present in inhalable particulate matter collected sampling time was 24 h for both sites. Prior to sampling, at two sites of São Paulo State (São Paulo and Piracicaba quartz fibre filters (20 × 25 cm, Millipore, USA) were cities). The results obtained in Piracicaba are the first baked in a muffle furnace at 650 °C for 6 h to eliminate reference of the determination of pesticides in particulate organics residues. In addition, filters were equilibrated matter collected in this site. In addition, the found pesticides at room temperature and weighed in a microbalance, levels were the basis to calculate the daily inhalation before and after the sampling, in order to estimate the exposure (DIE), the risk assessment, and the cancer risk concentration of particulate matter (PM). After sampling in all samples. and weighing, the filters were wrapped in aluminium foil and stored in a refrigerator at 5 °C until chemical analyses Experimental were performed. The meteorological data (ambient temperature, Site characterization and sampling precipitation, and wind speed) were collected from the database of the agrometeorological station at School of Piracicaba (PRB) is a city located in São Paulo State Agriculture (LEB, ESALQ, USP) for the campaigns in (Figure S1, Supplementary Information (SI) section). Its PRB.11 The meteorological data for SPA were collected population is 400 thousand and it is considered a rural area, from the climatological bulletin of the meteorological with over 94 thousand ha of planted area (approximately station at Institute of Astronomy, Geophysics and 70% of its total area), with the main crops being sugarcane Atmospheric Sciences (IAG, USP).12 and oranges.8 The most used pesticides in these crops are azoxystrobin, bifenthrin, carbofuran and λ-cyhalothrin. Reagent and chemicals The study site is localized within the School of Agriculture (University of São Paulo). Sugarcane crops surround the Environmental Protection Agency (EPA) organochlorine school and most of its area corresponds to experimental mix standard with α-hexachlorocyclohexane (α-HCH), crops. PM10 samples (n = 8) were collected in PRB in g-HCH, β-HCH, heptachlor, δ-HCH aldrin, heptachlor- Vol. 31, No. 6, 2020 Yera et al. 1319 epoxide, α-endosulfan, dichloro-diphenyldichloroethylene standard deviation (RSD, %) values for intraday precision (4,4’-DDE), dieldrin, endrin, dichloro- ranged from 0.19 (permethrin II) to 3.4% (heptachlor diphenyldichloroethane (4,4’-DDD), β-endosulfan, epoxide). LOD values ranged from 0.14 (δ-HCH) to dichloro-diphenyltrichloroethane (4,4’‑DDT), endrin 0.44 ng mL−1 (permethrin II) and LOQ values ranged from aldehyde and methoxychlor were acquired from Sigma- 0.63 (diazinon) to 1.5 ng mL−1 (permethrin II) (Table S1, Aldrich (St. Louis, USA). Malathion, parathion, sulfotep, SI section). The method performance for organochlorine fenthion, disulfoton, demeton-o, and bifenthrin standards pesticides was assessed by using surrogate standards were purchased from AccuStandard (New Haven, USA). TCMX and PCB103. These were added to blank samples, TCMX (2,4,5,6-tetrachloro-m-xylene) and PCB103 solvent blanks and samples before extraction. The average (2,2’,4,5’,6-pentachlorobiphenyl) were used as surrogate recoveries of TCMX and PCB103 in all samples ranged standards and were purchased from Ultra Scientific (North from 90 to 144% with RSD lower than 14%. Kingstown, USA). Acetonitrile were purchased from The pesticides identification and confirmation by Merck (Darmstadt, Germany). Ethyl acetate was supplied GC‑MS were performed according to criteria established by Macron (Center Valley, USA). All organic solvents by SANCO:14 (i) at least three diagnostic ions; (ii) retention used were spectroscopic and high-performance liquid time of the analyte in the sample should be similar to chromatography (HPLC) grade. standard within ± 0.20 min; (iii) signal-to-noise (S/N) of the A mix working solution containing all pesticides was least intense ion > 3:1.14 The qualifier ions used to pesticide prepared in ethyl acetate. Matrix-matching calibration identification are shown in the Table S1 (SI section). curves were prepared for each compound, adding varied volumes of the mix working solution in extracts of blank Quality assurance/quality control filters, as described in Nascimento et al.13 Each calibration curve is composed of seven levels of concentrations 0.5, All non-volumetric glassware, including the borosilicate 1.0, 2.5, 4.5, 6.5, 8.5, 10.5 ng mL−1.13 glass chamber, was cleaned with organic solvents (acetone and hexane) and then, baked in a muffle furnace at 450 °C for Extraction and sample analysis 4 h. After cleaning procedure, glassware blanks were tested, and no interfering peaks were observed. The instrument, The pesticides determination was carried out according reagent, method and field blanks were evaluated. The to method previously published elsewhere.13 Briefly, instrument blank consisted of GC-MS carrier gas analysis a 4.15 cm2 filter section was cut and transferred to a in order to detect the presence of interfering compounds microextraction device with borosilicate glass chamber and level of instrumental noise. The reagent blank was (Whatman Mini-UniPrep G2, USA). Then, 10 µL of the evaluated injecting extraction blanks containing only the surrogate standard TCMX and PCB103 with concentrations extraction solvent (ethyl acetate/acetonitrile mixture, 30:70) of 6.5 ng mL−1 were added to the particles in the filter. After into GC-MS. The method blank was assessed by extracting surrogate standard adsorption (2 h), 500 µL of a binary and further analysis of blank filters (without particles). The solvent mixture composed of ethyl acetate/acetonitrile field blank was assessed by analyzing a non-sampled filter. mixture (30:70) was added and the whole system was If any interfering peak are detected in the same retention sonicated during 23 min at 39 °C using an ultrasonic bath time of target pesticides, then the peak was discounted of (Symphony-VRW, Radnor, Pennsylvania, USA). Then, the original sample. extract was instantly filtered in the same device and injected into gas chromatograph coupled to mass spectrometer Inhalation exposure and risks assessment (GC-MS). The inhalation is an important source of atmospheric Analytical performance and identification criteria pesticides exposure. Following the methodology and the equations proposed by Coscollà and Yusà,1 the daily The analytical performance of the method was assessed inhalation exposure (DIE), the risk assessment and considering the following figures-of-merit: linear range, the cancer risk for three different populations: infants linearity, limit of detection (LOD), limit of quantification (6 months to 1.5 years), children (1.5 to 6 years) and adults (LOQ), instrumental precision (by mean of intraday and (> 12 years), were calculated.1 interday precision).13 All calibration curves presented The DIE (mg kg−1 day−1) refers to the amount of suitable determination coefficient values ranging from pesticides to which a given population is daily exposed. It 0.9903 (β-HCH) to 0.9985 (δ-HCH). The relative was determined using the equation 1. 1320 Occurrence of Pesticides Associated to Atmospheric Aerosols J. Braz. Chem. Soc.

(1) considering both sampling sites. For PRB samples, the compounds with the highest detection frequency (greater than 50%), taking into account values above the LOD, were where C (mg m−3) is the total concentration (gas + heptachlor and ethion, β-endosulfan, bifenthrin, permethrin I particulate phase) of each pesticide; IRinh (m3 h−1) is the and II, λ-cyhalothrin and demeton-o. For SPA samples, inhalation rate per hour, the IRinh applied was 8 m3 day−1 the pesticides with the highest detection frequencies were for infants, 10 m3 day−1 for children and 20 m3 day−1 for permethrin I and II, diazinon, β-endosulfan, bifenthrin and adults; ED is the exposure duration (h), for the three groups ethion (Tables S3 and S4, SI section). The concentrations of individuals were considered 24 h of exposure; BW is the ranged from 15 pg m−3 for tebuconazole to 160 pg m−3 for body weight (kg), which was considered to be 10 kg for ethion in PRB and from 15 pg m−3 (diazinon) to 166 pg m−3 infants, 15 kg for children and 70 kg for adults. (endrin aldehyde) in SPA. Figures 1 and 2 show the range The risk assessment was calculated using the hazard and average values of the concentration of pesticides for quotient (HQ) as a descriptor of the risk (equation 2). PRB and SPA, respectively.

(2) where HBRV is the health based reference values and it was calculated using the AOEL, defined as the acceptable operator exposure levels.1 HQ values higher than 1 mean that the populations are more exposed to the pesticides than the operator (person who works directly with the formulation). The cancer risk was calculated by the equation 3. It refers to compounds classified by the Environmental Protection Agency (EPA) as “likely to be carcinogenic to humans” and/or “possible human carcinogen” (diazinon, heptachlor, 4,4’-DDE, tebuconazole, permethrin I and permethrin II).15

Cancer risk = DIE × PF (3)

Potency factor (PF) is the factor that estimates the Figure 1. Concentrations of pesticides in the particulate phase at potency of the carcinogenicity. Lee et al.16 presented a Piracicaba (PRB). table with these values for some pesticides. In general, these values range from 10−3 to 10−1.16 Some values of PF for specific pesticides were not found, and 0.1 was assumed for all pesticides (situation with major potency).

Total concentration (C)

C is the sum of the concentrations in both gaseous and particulate phases. In the present study, only pesticides in particulate matter were determined. However, the corresponding gas phase pesticides concentrations were estimated using the octanol-air model (Koa) proposed by Harner and Bidleman17 (SI section).

Results and Discussion

Concentrations of pesticides in the particulate matter

Figure 2. Concentrations of pesticides in the particulate phase at São Twenty pesticides were detected and 14 were quantified, Paulo (SPA). Vol. 31, No. 6, 2020 Yera et al. 1321

Bifenthrin and permethrin are insecticides of the crop in PRB.10 In this site, the average concentration was pyrethroids group, and they are widely used in Brazil for 64 pg m−3. The highest and average concentrations found the application in plantations such as sugarcane, corn, in SPA were 54 and 37 pg m−3, respectively. Wang et al.24 coffee, soybean and bean.5 These five leguminous species found average concentration of 34.5 pg m−3 in the summer are among the ten most planted in São Paulo State,7 which of Vietnam. Heptachlor is rapidly converted to heptachlor- explains their high detection frequencies. epoxide mainly by biodegradation. Therefore, when the Bifenthrin is predominant in the particulate phase concentrations of heptachlor are greater than heptachlor- due to its low vapor pressure (1.8 × 10−5 Pa),18 which epoxide, it can indicate a recent use of the product according would explain the high detection frequency. Despite the to Secretary of Health of São Paulo State report (2003).25 In fact, its concentrations were below to LOQ in both sites. this work, the average heptachlor concentrations (64 pg m−3 Concentrations of bifenthrin over 83 pg m−3 and detection in PRB and 37 pg m−3 in SPA) exceeds the atmospheric −3 −3 frequency of 47% were found in PM10 samples in an area heptachlor-epoxide levels (26 pg m in PRB and 27 pg m of Valencia Region, Spain.19 in SPA) (Figures 1 and 2). Endosulfan (α and β) were largely used in Brazil to Ethion was the compound that presented the highest control insects populations in sugarcane, cocoa, coffee, concentrations in PRB and was found in 100% of the soybean, and cotton plantations. It was only totally banned samples, with average concentration of 82 pg m−3 and in Brazil in 2013.5 The maximum concentration found maximum concentration of 161 pg m−3 (Figure 1). For the for β-endosulfan in SPA was 96 pg m−3 and in PRB was samples collected in SPA, ethion was only detected in 53% 52 pg m−3. The α-endosulfan was not found in SPA samples, of the samples, and the highest concentrations were over while its average concentrations in the PRB samples are 25 pg m−3. In turn, diazinon is used in fruits and vegetables over 35 pg m−3 (Figures 1 and 2). crops and in the insects control in ornamental plants.10 Compounds like α‑ and β‑endosulfan present vapor In PRB it was not detected, while in SPA the detection pressures (0.0044 and 0.0040 Pa, respectively) suggesting frequency was at 93% of the samples and the maximum that they are mostly in the gas phase.20 Few works concentration reached 31 pg m−3. In other urban areas, as determined the concentrations of these compounds only in in the case of Valencia (Spain), average concentrations of the particulate phase. Chrysikou et al.21 and Coscollà et al.22 diazinon over 30 pg m−3 are reported,26 a result similar to did not detect in PM2.5 collected in rural and urban cities in that determined in this work. Europe. However, values of 29 pg m−3 are reported in the Even though 34 pesticides are investigated in the gas phase in a remote area in southern Brazil.23 samples collected in SPA and PRB, only 14 pesticides Concentrations of α-endosulfan are generally greater were found in most of the samples of both sites. In PRB, than β-endosulfan because the latter is more unstable, it eight pesticides presented a high frequency of detection, is more prone to undergo degradation or transformation while in SPA only six of them presented a high frequency into its other isomer. In addition, the losses due to wet of detection (Tables S3 and S4, SI section). For PRB, the depositions are greater for β-endosulfan because it has a high pesticides frequencies and concentrations may be due higher solubility.23 to the wide use of a large amount of pesticides in the area More than 95% of the endosulfan formulation corresponds nearby the sampling site. to its α and β isomers.20 The α:β ratio varies with the Figure 3 shows the variation of the sum of de manufacturer but it is generally in the range of 2:1 to 7:3. concentrations of pesticides (ΣCpesticides) in the particulate Elevated α:β ratio represents an aged signature scenario for phase and some meteorological parameters at PRB site. The 23 −3 these pesticides. In this work, the ratio for PRB, considering highest value found for ΣCpesticides was 484 pg m , while the −3 −3 the average of all the samples (35 pg m for α-endosulfan lowest value was 69 pg m . Concentrations of PM10 ranged and 38 pg m−3 for β-endosulfan), was 0.9, which suggests a from 26 to 120 µg m−3.27 Days that presented high and low recent application of this pesticide nearby the sampling area. values of concentrations of PM10 presented similar values of

Heptachlor is highly toxic to human health, banned in ΣCpesticides. The rainfall occurred in only 2 days of sampling, in −3 the countries of the European Union. In Brazil, up to 2007, which the ΣCpesticides were 379 and 206 pg m , respectively. its use was only allowed for wood preservation. But since The sample collected in the warmest day (PRB 7, 27 °C) 2016, it is prohibited for any use in Brazil.5 In PRB, it and with highest wind speed (12 km h−1) presented the lower −3 was detected in all samples, and in a low frequency (40%) value of ΣCpesticides (69 pg m ). as well as in the samples of SPA, which were collected For SPA samples, the lowest ΣCpesticides was over in 2017 (after its total prohibition). This pesticide is 31 pg m−3 and the highest was 372 pg m−3. Concentrations −3 intensely used for sugarcane pests’ control, a predominant of PM2.5 ranged from 6 to 79 µg m . On the sampling days, 1322 Occurrence of Pesticides Associated to Atmospheric Aerosols J. Braz. Chem. Soc.

Figure 3. Sum of the concentrations of all pesticides (ΣCpesticides) present in particulate phase for each sample and meteorological parameter at PRB site.

ΣCpesticides are represented by green bars (left axis), whereas concentrations of PM10, rainfall, temperature and wind speed are represented as line and point (right axis).

the temperature ranged from 14 to 21 °C. The rainfall, in the particulate phase, concentrations of PM10 and nevertheless, occurred in 6 sampling days, reaching the meteorological parameter. In general, the correlations maximum of 32 mm (SPA 75, Figure 4). The sample with between pesticides depends on the chemical class −3 the highest ΣCpesticides was SPA 68 (372 pg m ), which also (organochlorines, organophosphorus, pyrethroids and presented high concentration of particulate matter PM2.5 triazole) and application frequency. The greatest correlation (30 µg m−3), the highest ambient temperature (21 °C) and no found (0.95) was for demeton-o and malathion, both rainfall episode; it is expected that pollutants are normally organophosphorus pesticides. They are used as insecticide eliminated from the atmosphere by wet deposition. In this in sugarcane crops and have the same mechanism of day, pesticides input in the atmosphere was maximized action, so their atmospheric reactions or degradation since the only atmospheric removal mechanisms were dry could be similar. High correlation was also found between deposition and degradation. permethrin I and permethrin II. These isomers are the Pearson analysis for the PRB samples showed the components of the commercial formulation and have the correlations between the concentrations of pesticides same chemical behavior in the atmosphere.

Figure 4. Sum of the concentrations of all pesticides (ΣCpesticides) present in particulate phase for each sample and meteorological parameter at SPA site.

ΣCpesticides are represented by green bars (left axis), whereas concentrations of PM10, rainfall, temperature and wind speed are represented as line and point (right axis). Vol. 31, No. 6, 2020 Yera et al. 1323

Negative correlation was found for heptachlor and comparing the vapor pressure of these pesticides, it and heptachlor epoxide (−0.74). It is expected since was observed that compounds like demeton-o, diazinon, heptachlor epoxide is the main product of the heptachlor heptachlor, malathion, heptachlor-epoxide, 4,4’‑DDE, degradation.25 It is reasonable, that as the concentration α-endosulfan, β-endosulfan and endrin aldehyde, are prone of heptachlor decreases, the concentration of heptachlor to be found in the gaseous phase. Ethion, permethrin I and epoxide deceases. This hypothesis is also demonstrated permethrin II are distributed in both phases. Tebuconazole with the fact that, for the compounds that present high and λ-cyhalothrin are predominantly found in the particulate correlation with heptachlor (demeton-o: 0.75, malathion: phase (Tables S5 and S6, SI section). 0.84, α-endosulfan: 0.83 and endrin aldehyde: 0.74), For compounds with very small Φ (10−4 to 10−2) the total these correlations are negative with heptachlor epoxide concentrations were higher than compounds with high Φ (demeton-o: −0.53, malathion: −0.56, α-endosulfan: −0.74 (Tables S5 and S6, SI section). This is because the most and endrin aldehyde: −0.57). Similarly, the β-endosulfan part of these compounds is in the gaseous phase, so that, presented positive correlation with heptachlor epoxide practically the whole amount of them was in the gas phase

(0.60) and negative with heptachlor (−0.58). and they were estimated through the Koa model. α-Endosulfan and β-endosulfan are the components DIE values for three different population groups (adults, of the commercial endosulfan formulation, being children and infants) were calculated for both sampling α-endosulfan found in higher concentration than its isomer. sites considering the average (Table S7, SI section) and the α-Endosulfan can be transformed to β-endosulfan in the maximum concentrations (Table 1). The DIE values range environment,23 which could explain the negative correlation from 5 × 10−9 mg kg−1 day−1 for tebuconazole in adults to found for these compounds (−0.62). 1 × 10−4 mg kg−1 day−1 for heptachlor in infant at PRB, Low correlations were found between concentrations while in SPA, the values ranged from 6 × 10−9 mg kg−1 day−1 of pesticides and meteorological parameters. The only for tebuconazole in adults to 9 × 10−5 mg kg−1 day−1 for pesticide that presented a moderate negative correlation heptachlor in infants. In general, it is expected that the was ethion with wind speed (−0.69). A general analysis highest DIE values are attributed to the most vulnerable of the correlations between the individual pesticides and population (infants) because they have lower body weight.1 the meteorological conditions in this rural area indicates For demeton-o and malathion the DIE maximum values that the concentrations variation depends more on the in infants at PRB were 7 × 10−6 and 1 × 10−6 mg kg−1 day−1, physical and chemical properties of the compounds than respectively, and 4 × 10−6 and 9 × 10−6 mg kg−1 day−1 at on meteorological conditions. SPA. These results are slightly higher than that reported For the SPA samples, the Pearson analysis does not by Nascimento et al.13 in Salvador, northeastern Brazil, provide much information. The correlations between 4 × 10−8 and 5 × 10−8 mg kg−1 day−1 in infants for demeton-o pesticides concentration are generally low. The pesticide and malathion, respectively.13 Low DIE were also found with the highest number of positive and high correlation for malathion in Valencia region in different years, was diazinon (4,4’-DDE: 0.72, ethion: 0.83, and endrin 2 × 10−7 mg kg−1 day−1 in 2010 and 6 × 10−7 mg kg−1 day−1 aldehyde: 0.80). These correlations are difficult to explain in 2014.26,29 since these pesticides do not have similar physical and The hazard quotient, HQAOEL, gives us a measure of chemical properties. Regarding the temperature, moderate how close the daily inhalation value is to the highest value positive correlations were found with diazinon (0.60) that an operator can be exposed. Values greater than 1 and ethion (0.71). The increase in temperature favors the indicate a risk to the human health.1 Table 1 compares the compounds volatilization from the ground, plants and results obtained of HQAOEL for the maximum concentrations other surfaces.28 of pesticides in adults, children, and infants; Table S7

(SI section) shows the HQAOEL values for the average Risk assessment concentrations of pesticides.

In PRB the highest values of HQAOEL were found in The fraction of the pesticides in the particulate phase (Φ) infants (α-endosulfan: 4 × 10−3, β-endosulfan: 1 × 10−3, and was calculated to estimate the concentration in the gaseous ethion: 3 × 10−4), while for SPA the highest values were for −3 −3 phase and the total concentration (Ctotal) of the pesticides. diazinon: 7 × 10 , β-endosulfan: 3 × 10 and malathion: Compounds with vapor pressure lower than 10−5 can be 3 × 10−4. Values reported29 in Valencia for tebuconazole mostly found in the particulate phase; between 10−5 to 10−2 (6 × 10−3) in infants were higher than the values found for can be distributed in both phases and higher than 10−2 can be PRB (5 × 10−7) and for SPA (6 × 10−7). All the values found mostly found in the gaseous phase.26 Following this approach in this work are below 1.0, indicating that there is no risk to 1324 Occurrence of Pesticides Associated to Atmospheric Aerosols J. Braz. Chem. Soc.

Table 1. Maximum concentrations in particulate and gaseous phase (Cmax), daily inhalation exposure (DIE) and hazard quotient (HQAOEL) of the most detected pesticides in both sampling sites for infants, children and adults

−3 Cmax / (pg m ) Infants Children Adults PRB SPA PRB SPA PRB SPA Pesticide PRB SPA DIE / DIE / DIE / DIE / DIE / DIE / HQ HQ HQ HQ HQ HQ (mg kg−1 day−1) AOEL (mg kg−1 day−1) AOEL (mg kg−1 day−1) AOEL (mg kg−1 day−1) AOEL (mg kg−1 day−1) AOEL (mg kg−1 day−1) AOEL Demeton-o 8262 5090 7 × 10−6 − 4 × 10−6 − 6 × 10−6 − 3 × 10−6 − 2 × 10−6 − 2 × 10−6 − Diazinon − 1711 − 1 × 10−6 7 × 10−3 − 1 × 10−6 6 × 10−3 − − 5 × 10−7 2 × 10−3 Heptachlor 175299 110000 1 × 10−4 − 9 × 10−5 − 1 × 10−4 − 7 × 10−5 − 5 × 10−5 − 3 × 10−5 − Malathion 1700 10600 1 × 10−6 5 × 10−5 9 × 10−6 3 × 10−4 2 × 10−6 4 × 10−5 7 × 10−6 2 × 10−4 5 × 10−7 9 × 10−6 3 × 10−6 1 × 10−4 Heptachlor-epoxide 11200 21000 9 × 10−6 − 2 × 10−5 − 8 × 10−6 − 1 × 10−5 − 3 × 10−6 − 6 × 10−6 −

α-Endosulfan 8790 − 7 × 10−6 4 × 10−3 − 6 × 10−6 3 × 10−3 - − 3 × 10−6 7 × 10−4 − − 4,4’-DDE − 559 − 5 × 10−7 − − 4 × 10−7 − − − 2 × 10−7 − Ethion 616 115 5 × 10−7 3 × 10−4 9 × 10−8 6 × 10−5 4 × 10−7 3 × 10−4 8 × 10−8 5 × 10−5 2 × 10−7 8 × 10−5 3 × 10−8 2 × 10−5

β-Endosulfan 2480 7679 2 × 10−6 1 × 10−3 6 × 10−6 3 × 10−3 2 × 10−6 8 × 10−4 5 × 10−6 3 × 10−3 7 × 10−7 2 × 10−4 2 × 10−6 1 × 10−3 Endrin aldehyde 2861 5191 2 × 10−6 − 4 × 10−6 − 2 × 10−6 − 4 × 10−6 − 8 × 10−7 − 2 × 10−6 − Tebuconazole 18 23 2 × 10−8 5 × 10−7 2 × 10−8 6 × 10−7 1 × 10−8 4 × 10−7 2 × 10−8 5 × 10−7 5 × 10−9 2 × 10−7 7 × 10−9 2 × 10−7

λ-Cyhalothrin 30 47 2 × 10−8 8 × 10−7 4 × 10−8 1 × 10−6 2 × 10−8 7 × 10−7 3 × 10−8 1 × 10−6 9 × 10−9 5 × 10−4 1 × 10−8 5 × 10−7 Permethrin I 103 319 8 × 10−8 2 × 10−6 3 × 10−7 5 × 10−6 7 × 10−8 1 × 10−6 2 × 10−7 4 × 10−6 3 × 10−8 5 × 10−7 9 × 10−8 2 ×10−6 Permethrin II 235 426 1 × 10−7 4 × 10−6 3 × 10−7 7 × 10−6 2 × 10−7 3 × 10−6 3 × 10−7 6 × 10−6 7 × 10−8 1 × 10−6 1 × 10−7 2 ×10−6 PRB: Piracicaba; SPA: São Paulo; −: compound not detected; 4,4’-DDE: dichloro-diphenyldichloroethylene. human health by inhalation exposure to these compounds. for PRB and SPA. Table 2 shows the cancer classification proposed by Pesticides classified by the EPA as “likely to be the EPA, and the results obtained in this study for the carcinogenic to humans” and “possible human carcinogen” maximum values of DIE in adults, children and infants, are interesting because high concentrations of these

Table 2. Cancer classification and cancer risk for pesticides in Piracicaba (PRB) and São Paulo (SPA)

Cancer risk Pesticide Classification Infants Children Adults PRB SPA PRB SPA PRB SPA Demeton-o not carcinogenica 7 × 10−7 5 × 10−7 6 × 10−7 3 × 10−7 2 × 10−7 2 × 10−7 Diazinon likely carcinogenicb − 1 × 10−7 − 1 × 10−7 − 5 × 10−8 Heptachlor possiblec 1 × 10−5 9 × 10−6 1 × 10−5 7 × 10−6 5 × 10−6 3 × 10−6 Malathion not evidenced 1 × 10−7 9 × 10−7 1 × 10−7 7 × 10−7 5 × 10−8 3 × 10−7 Heptachlor-epoxide − 9 × 10−7 2 × 10−6 8 × 10−7 1 × 10−6 3 × 10−7 6 × 10−7 α-Endosulfan not carcinogenica 7 × 10−7 − 6 × 10−7 − 3 × 10−7 − 4,4’-DDE likely carcinogenicb − 5 × 10−8 − 4 × 10−8 − 2 × 10−8 Ethion evidencee 5 × 10−8 9 × 10−9 4 × 10−8 2 × 10−9 2 × 10−8 3 × 10−9 β-Endosulfan not carcinogenica 2 × 10−7 6 × 10−7 2 × 10−7 5 × 10−7 7 × 10−8 2 × 10−7 Endrin aldehyde − 2 × 10−7 4 × 10−7 2 × 10−7 4 × 10−7 8 × 10−8 2 × 10−7 Tebuconazole possiblec 2 × 10−9 2 × 10−9 1 × 10−9 2 × 10−9 5 × 10−10 7 × 10−10 λ-Cyhalothrin − 2 × 10−9 4 × 10−9 2 × 10−9 3 × 10−9 9 × 10−10 1 × 10−9 Permethrin I likely carcinogenicb 8 × 10−9 3 × 10−8 7 × 10−9 2 × 10−8 3 × 10−9 9 × 10−9 Permethrin II likely carcinogenicb 2 × 10−8 3 × 10−8 2 × 10−8 3 × 10−8 7 × 10−9 1 × 10−8 aNot likely to be carcinogenic to humans; blikely to be carcinogenic to humans; cpossible human carcinogenic; dsuggestive evidence of carcinogenicity, but not sufficient to assess human carcinogenic potential; eevidence of no carcinogenicity for humans.30 −: compound not detected; 4,4’-DDE: dichloro- diphenyldichloroethylene. The cancer risk was calculated for the compounds with the highest detection frequency. Vol. 31, No. 6, 2020 Yera et al. 1325 compounds increase the risk of cancer in the population.30 FAPESB (projects 8164/2014 and 2159/2012), Financiadora The maximum value acceptable for the cancer risk is de Projetos e Estudos (01.14.0215.00) and FAPESP (project 1 × 10−6.26 Higher values can be interpreted as excess cancer 2017/20826-1). J. B. A., G. O. R. and P. C. V. thank CNPq risk for a given population. for their research fellowships. For the compounds classified as: likely carcinogenic, possible carcinogenic, no evidence and evidence of References carcinogenicity, the highest values observed in infants, children and adults, respectively, for heptachlor were: 1. Coscollà, C.; Yusà, V.; Comprehensive Analytical Chemistry, 1 × 10−5, 1 × 10−5 and 5 × 10−6 at PRB, and 9 × 10−6, 7 × 10−6 vol. 73.; Elsevier: London, UK, 2016. and 3 × 10−6 at SPA. These values exceeded the maximum 2. Villiot, A.; Chrétien, E.; Drab-Sommesous, E.; Rivière, E.; allowable value, indicating cancer risk for these populations Chakir, A.; Roth, E.; Atmos. Environ. 2018, 174, 82. at both sampling sites. Another pesticide, which exceeded 3. Viel, J. F.; Floret, N.; Deconinck, E.; Focant, J. F.; De Pauw, this limit, was heptachlor epoxide, in infants and children E.; Cahn, J. Y.; Environ. Int. 2011, 37, 449. at SPA site. As mentioned above, the carcinogenic activity 4. Casida, J. E.; Durkin, K. A.; Annu. Rev. Entomol. 2013, 58, 99. for this compound was not reported (Table 2). 5. http://portal.anvisa.gov.br/registros-e- autorizacoes/agrotoxicos/produtos/monografia- Conclusions de-agrotoxicos/autorizadas, accessed in January 2020. 6. http://revistapesquisa.fapesp.br/wp-content/ In this work, the concentration of pesticides in the uploads/2018/09/018-027_CAPA-Agrotóxicos_271.pdf, atmosphere of São Paulo and Piracicaba cities (sugarcane accessed in January 2020. plantation site) was determined and the hazard quotients 7. Bombardi, L. M.; Geografia de Uso de Agrotóxicos no Brasil and cancer risk caused by inhalation of these compounds e Conexões com a União Europeia, vol. 1, 1st ed.; USP: São were calculated. Twenty-three samples were analyzed, and Paulo, Brazil, 2017. 34 pesticides associated to atmospheric particulate matter 8. https://cidades.ibge.gov.br/brasil/sp/piracicaba/panorama,

(PM2.5 and PM10) were investigated by GC-MS. Fourteen accessed on April 24, 2019. compounds including organochlorines, organophosphorus 9. https://cidades.ibge.gov.br/brasil/sp/sao-paulo/panorama, and pyrethroids pesticides, were found in these samples. accessed on April 24, 2019. Among them, heptachlor and ethion were found in all 10. http://agrofit.agricultura.gov.br/agrofit_cons/principal_agrofit_ samples collected in Piracicaba and permethrin I and II in cons, accessed in January 2020. São Paulo, as well. 11. http://www.leb.esalq.usp.br/leb/exceldados/DCE2008.TXT, The values for DIE were calculated for infants, children accessed in January 2020. and adults and they do not suggest inhalation hazard, i.e., 12. http://www.estacao.iag.usp.br/Boletins/2017.pdf, accessed in respiratory problems, for this population. Despite this, January 2020. heptachlor, compound classified as possible carcinogenic 13. Nascimento, M. M.; da Rocha, G. O.; de Andrade, J. B.; for humans, presented high cancer risk values for all Microchem. J. 2018, 139, 424. population, suggesting that the exposure to this compound 14. European Commission (EC); SANCO/12571/2013: Guidance can be dangerous. Document on Analytical Quality Control and Validation This work could be a good contribution to better Procedures for Pesticides Residues Analysis in Food and understand the role of pesticides associated to inhalable Feed; European Commission Health & Consumer Protection atmospheric particulate matter, and the hazard quotients Directorate-General and Safety of the Food Chain Chemicals, and cancer risk caused by inhalation of these compounds. Contaminants, and Pesticides, 2013; available at http:// www.eurl-pesticides.eu/library/docs/allcrl/AqcGuidance_ Supplementary Information Sanco_2013_12571.pdf, accessed in January 2020. 15. EPA; Guidelines for Carcinogen Risk Assessment; 2005, Supplementary data are available free of charge at available at https://www.epa.gov/sites/production/files/2013-09/ http://jbcs.sbq.org.br as PDF file. documents/cancer_guidelines_final_3-25-05.pdf, accessed in January 2020. Acknowledgments 16. Lee, S.; McLaughlin, R.; Harnly, M.; Gunier, R.; Kreutzer, R.; Environ. Health Perspect. 2002, 110, 1175. The authors thank CNPq (projects 465497/2014-4, 17. Harner, T.; Bidleman, T. F.; Environ. Sci. Technol. 1998, 32, 133711/2017-0), CAPES (project 88887.136374/2017‑00), 1494. 1326 Occurrence of Pesticides Associated to Atmospheric Aerosols J. Braz. Chem. Soc.

18. https://sitem.herts.ac.uk/aeru/iupac/Reports/78.htm, accessed 26. Yusà, V.; Coscollà, C.; Millet, M.; Atmos. Environ. 2014, 96, in January 2020. 322. 19. Hart, E.; Coscollà, C.; Pastor, A.; Yusà, V.; Atmos. Environ. 27. Vasconcellos, P. C.; Souza, D. Z.; Sanchez-Ccoyllo, O.; 2012, 62, 118. Bustillos, J. O.; Lee, H.; Santos, F. C.; Nascimento, K. H.; 20. Weber, J.; Halsall, C. J.; Muir, D.; Teixeira, C.; Small, J.; Araújo, M. P.; Saarnio, K.; Teinilä, K.; Hillamo, R.; Sci. Total Solomon, K.; Hermanson, M.; Hung, H.; Bidleman, T.; Sci. Environ. 2010, 408, 5836. Total Environ. 2010, 408, 2966. 28. Gevao, B.; Porcelli, M.; Rajagopalan, S.; Krishnan, D.; 21. Chrysikou, L. P.; Gemenetzis, P. G.; Samara, C. A.; Atmos. Martinez-Guijarro, K.; Alshemmari, H.; Bahloul, M.; Zafar, Environ. 2009, 43, 290. J.; Sci. Total Environ. 2018, 622-623, 1621. 22. Coscollà, C.; López, A.; Yahyaoui, A.; Colin, P.; Robin, C.; 29. López, A.; Yusà, V.; Muñoz, A.; Vera, T.; Borràs, E.; Ródenas, Poinsignon, Q.; Yusà, V.; Sci. Total Environ. 2017, 584-585, M.; Coscollà, C.; Sci. Total Environ. 2017, 574, 724. 856. 30. http://npic.orst.edu/chemicals_evaluated.pdf, accessed in 23. Meire, R. O.; Khairy, M.; Targino, A. C.; Galvão, P. M. A.; January 2020. Torres, J. P. M.; Malm, O.; Lohmann, R.; Chemosphere 2016, 144, 2175. Submitted: October 30, 2019 24. Wang, W.; Wang, Y.; Zhang, R.; Wang, S.; Wei, C.; Chaemfa, Published online: February 3, 2020 C.; Li, J.; Zhang, G.; Yu, K.; Sci. Total Environ. 2016, 542, 777. 25. http://acpo.org.br/paulinia/paulinia.pdf, accessed in January 2020.

This is an open-access article distributed under the terms of the Creative Commons Attribution License.